Anti-SARM1 antibody [EPR24834-80]

15 product images

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  Multiplex immunohistochemistry - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse colon tissue staining GSDMC2 + GSDMC3 with Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] ab229896 at a 1:2000 (0.25 ug/ml) dilution; SARM1 with ab309195 at 1:500 (0.964 ug/ml) dilution and MUC2 with Anti-MUC2 antibody [EPR23479-47] ab272692 at 1:2000 (0.252 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

  Panel A: merged staining of anti-SARM1 (green; Opal^™520), anti-MUC2 (gray; Opal^™570) and anti-GSDMC2/3 (magenta; Opal^™690)on mouse colon.
  
  Panel B: anti-SARM1 staining myenteric nerve plexus in mouse colon.
  
  Panel C: anti-MUC2 staining goblet cells in mouse colon.
  
  Panel D: anti-GSDMC2/3 staining epithelium in mouse colon.
  
  Nuclear DNA was labeled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of ab309195, Anti-MUC2 antibody [EPR23479-47] ab272692 and Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] ab229896 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

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  Immunoprecipitation - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  SARM1 was immunoprecipitated from 0.35 mg Rat brain tissue lysate with ab309195 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309195 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  Lane 1: Rat brain tissue lysate
  Lane 2: ab309195 IP in Rat brain tissue lysate
  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab309195 in rat brain tissue lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 10 seconds

  All lanes: Immunoprecipitation - Anti-SARM1 antibody [EPR24834-80] (ab309195) at 1/30 dilution

  All lanes: Rat brain tissue lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 10s

• 

  Immunoprecipitation - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  SARM1 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab309195 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309195 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  Lane 1: Mouse brain tissue lysate
  Lane 2: ab309195 IP in Mouse brain tissue lysate
  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab309195 in mouse brain tissue lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 10 seconds

  All lanes: Immunoprecipitation - Anti-SARM1 antibody [EPR24834-80] (ab309195) at 1/30 dilution

  All lanes: Mouse brain tissue lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 10s

• 

  Immunoprecipitation - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  SARM1 was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate with ab309195 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309195 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  Lane 1: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
  Lane 2: ab309195 IP in SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab309195 in SH-SY5Y whole cell lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 10 seconds

  All lanes: Immunoprecipitation - Anti-SARM1 antibody [EPR24834-80] (ab309195) at 1/30 dilution

  All lanes: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 10s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SARM1 with ab309195 at 1/500 (0.964 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse cerebrum (PMID: 21555464). The section was incubated with ab309195 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling SARM1 with ab309195 at 1/500 (0.964 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat cerebrum. The section was incubated with ab309195 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling SARM1 with ab309195 at 1/500 (0.964 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control: No staining on mouse cardiac muscle. The section was incubated with ab309195 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• 

  Immunohistochemistry (Frozen sections) - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat heart (fresh) tissue labeling SARM1 with ab309195 at 1/50 (9.64 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control: confocal image showing no staining on rat heart (PMID: 21555464). The nuclear counterstain was DAPI (Blue). The section was incubated with ab309195 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue).
  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse heart (fresh) tissue labeling SARM1 with ab309195 at 1/50 (9.64 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control: confocal image showing no staining on mouse heart (PMID: 21555464). The nuclear counterstain was DAPI (Blue). The section was incubated with ab309195 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue).
  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh) tissue labeling SARM1 with ab309195 at 1/50 (9.64 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on rat hippocampus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309195 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue).
  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh) tissue labeling SARM1 with ab309195 at 1/50 (9.64 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on mouse hippocampus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309195 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue).
  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

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  Immunocytochemistry/ Immunofluorescence - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling SARM1 with ab309195 at 1/50 (9.64 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in SH-SY5Y cell line, and showing no staining in HaCaTcell line.Negative control: HaCaT.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

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  Western blot - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  
  Exposure time: 180 seconds

  All lanes: Western blot - Anti-SARM1 antibody [EPR24834-80] (ab309195) at 1/1000 dilution

  Lane 1: Human brain tissue lysate at 20 µg

  Lane 2: Rat spinal cord tissue lysate at 20 µg

  Lane 3: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

  Secondary

  Lane 1: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Lanes 2 - 3: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

  Observed band size: 70 kDa

  Exposure time: 180s

• 

  Western blot - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  Negative control: HaCaT, EL4, mouse heart (PMID:21555464).
  
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200, 000 dilution.
  Exposure time: 180 seconds

  All lanes: Western blot - Anti-SARM1 antibody [EPR24834-80] (ab309195) at 1/1000 dilution

  Lane 1: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HaCaT (human skin keratinocyte) whole cell lysate at 20 µg

  Lane 3: EL4 (mouse lymphoma t lymphocyte) whole cell lysate at 20 µg

  Lane 4: Mouse brain tissue lysate at 20 µg

  Lane 5: Mouse heart tissue lysate at 20 µg

  Lane 6: Rat brain tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

  Observed band size: 70 kDa

  Exposure time: 180s

• 

  Western blot - Anti-SARM1 antibody [EPR24834-80] (ab309195)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  Wild-type and KO lysates were kindly provided by Dr. Xiaodong Wang, NIBS.
  
  Performed under reducing conditions.
  
  In Western blot, ab309195 was shown to bind specifically to SARM1. Target of interest was observed at 70 kDa in wild-type mouse brain tissue lysate (lane 1) with no signal observed at this size in SARM1 knockout mouse brain tissue lysate (lane 2).
  
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200,000 dilution.
  Exposure time: 180 seconds

  All lanes: Western blot - Anti-SARM1 antibody [EPR24834-80] (ab309195) at 1/1000 dilution

  Lane 1: Wild-type mouse brain tissue lysate at 20 µg

  Lane 2: SARM1 knockout mouse brain tissue lysate at 20 µg

  Lane 3: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

  Lane 4: EL4 (mouse lymphoma t lymphocyte) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

  Performed under reducing conditions.

  Observed band size: 70 kDa

  Exposure time: 180s